Isolation of a cytochrome P-450 structural gene from Saccharomyces cerevisiae.
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Kalb VF, Loper JC, Dey CR, Woods CW, Sutter TR
Isolation of a cytochrome P-450 structural gene from Saccharomyces cerevisiae.
Gene. 1986;45(3):237-45.
- PubMed ID
- 3542713 [ View in PubMed]
- Abstract
We have transformed a Saccharomyces cerevisiae host with an S. cerevisiae genomic library contained in the shuttle vector YEp24 and screened the resultant transformants for resistance to ketoconazole (Kc), an inhibitor of the cytochrome P-450 (P-450) enzyme lanosterol 14 alpha-demethylase. Two plasmids were isolated which transformed yeast to both increased resistance to Kc and increased levels of total P-450. Hybrid-selection and immunoprecipitation experiments showed that these plasmids, pVK1 and pVK2, contained the structural gene for an S. cerevisiae P-450. This conclusion was confirmed by the nucleotide sequence of a portion of pVK2, which revealed an open reading frame encoding a characteristic P-450 heme-binding region.